Anti-viral triaza compounds and compositions

ABSTRACT

The invention relates to a family of new synthetic triamine compounds which can be used in antiviral pharmaceutical compositions.

This application is a 371 of PCT/US96/02132 filed Feb. 16, 1996 which isa continuation-in-part of U.S. application 08/392,550 filed Feb. 17,1995 now issued U.S. Pat. No. 5,663,161.

The invention relates to a family of new synthetic triamine compoundswhich can be used in pharmaceutical compositions such as antivirals.

BACKGROUND OF THE INVENTION

The number of biologically active compounds useful as antivirals is verylimited. Particularly limited are anti-HIV agents. Currently approvedanti-HIV drugs include nucleoside analogs such as AZT. Unfortunately,recent results of a European trial of AZT in asymptomatic HIV-infectedpersons showed that administration of the drug caused no difference insurvival after three years. In the face of slow progress on thedevelopment of an effective AIDS vaccine, a National Task Force on AIDSDrug Development has been established to explore new approaches to AIDSchemotherapy.

It is unlikely that a cure for AIDS will be discovered because HIVintegrates itself into the host's genome and the main strategy to combatthe disease is to keep the HIV virus from proliferating. At least 16steps in the HIV life cycle have been identified as possible points fortherapeutic intervention, yet all of the anti-HIV drugs licensed in theU.S. so far (AZT, ddI and ddC) are nucleoside inhibitors of HIV reversetranscriptase (RT). Rapid mutation of the virus presents a key challengeto antiretroviral drug therapy and AZT-resistant strains of HIV appearin patients after prolonged treatment. Non-nucleoside RT inhibitors arecurrently under investigation, but it is expected that combinations ofdrugs operating by different mechanisms will combat viral resistancemost successfully. Hence, there is an urgent search for new drugs actingat different stages in the HIV life cycle. More recently discoveredanti-HIV agents include certain bicyclams and quinolines such asquinoline-4-amine. The mechanism of action of the quinoline compound isunknown, but the bicyclams are reported to be inhibitors of HIVuncoating.

Another type of compound, the triaza macrocycles, have been usedprimarily for metal complexation. Previously, no biological activity hasbeen suggested for these compounds.

SUMMARY OF THE INVENTION

The present invention provides a family of triaza compounds which showantiviral properties for use against a number of viruses including HIV-1and HIV-2.

The basic structure of the compounds is represented by formula I:

wherein

W represents a bridge carbon which is additionally bonded to at leastone polar or non-polar side group substituent selected from the groupconsisting of double-bonded carbon, double bonded oxygen, hydroxyl,alkyl of about one to 10 carbons, alkoxy of about one to 10 carbons,aryl of about 7 to 10 carbons, a halogen, methyl halogen, methylenehalide, epoxide (or oxirane), acyl, CH₂OH and hydrogen; halogen is F,Cl, I or Br; halide is F₂, Cl₂, I₂ or Br₂.

X and Y independently represent an aromatic group, an alkyl group, asulfonyl group or a carbonyl group said aromatic group selected from thegroup consisting of Ar, Ar sulfonyl, Ar carboxy and Ar alkyl where Ar isan aromatic cyclic or heterocyclic ring having from five to sevenmembers. The alkyl group which may be present for X and Y or as asubstituent on Ar has from one to ten carbons. X and Y are not both analkyl group. Preferably at least one of X or Y is an aromatic group.

Z represents a group listed for X and Y or a fused aryl moiety; saidaryl moiety having from seven to ten carbons. Z may also representhydrogen.

a and d independently represent a number from zero to 10; b and cindependently represent a number from one to 10; and e represents anumber from zero to three; and preferably, a+d+e≧1. Formula I representsa cyclic or acyclic structure and contains sufficient hydrogens for astable molecule. Moieties for C_(a) and C_(d) can include double bondsparticularly when the structure for formula I is acyclic.

In one preferred embodiment, W is ethene, X and Y are tosyl, Z is benzylor —COR², a, d, and e are one, and b and c are three.

The compounds of formula I have antiinfective activity and have a rangeof uses including as a pharmaceutical agent for use in the preventionand chemoprophylaxis of viral illnesses. The compounds can also be usedas an antiinfective or antiseptic as a coating or additive on articlessuch as medical devices, contraceptives, dressings, and in blood productpreparations and similar biologicals.

A method of inhibiting a virus comprises contacting the virus, avirus-containing milieu, a virus-infectable cell or a virus-infectedcell with a virus-inhibiting amount of the compound of formula I.

DETAILED DESCRIPTION

The compounds are characterized as having at least three nitrogen atoms(amine sites) linked by at least two alkylene bridges or linking groupsto form triamines. The alkylene bridge linking groups are preferablyalkanes containing from one to ten carbons.

The triamines may be formed into a triazamacrocycle by a third alkylenebridge which is preferably alkane having from one to ten carbonsconnecting the two end nitrogens of the triamine compound.

The alkylene bridges linking the nitrogen atoms can additionally includearomatic or non-aromatic rings fused to the alkylene bridge. Bridgescontaining fused rings and linking two nitrogens of the triaminestructure may be exemplified by the following:

The alkylene bridges are preferably —(CH₂)₃—.

The bridge carbon (designated W) of the third alkylene bridge may befunctionalized with (i.e., bonded to) a side substituent which is apolar group.

Representative groups for W include

(R and R¹ are alkyl of 1-10 carbons),

(halo is F, Cl, Br or I). W may also be unfunctionalized, i.e., bondedto hydrogen. W is preferably

X and Y are independently an aromatic, alkyl, sulfonyl or carbonyl groupor hydrogen. Representative aromatic groups include five or six memberedrings which may have heteroatoms of nitrogen, oxygen or sulfur. Therings include, for example, phenyl, pyrrolyl, furanyl, thiophenyl,pyridyl, thiazoyl, etc. The aromatic group (Ar) for X and Y may besubstituted with a hydrophilic group. Preferably the Ar is substitutedwith NO, NO₂, NH₂, NHR, NHR₂, OH, OR, SH, SR, SOR, SO₃R, halo,C(halogen)₃

where R is alkyl of one to 10 carbons; R is preferably alkyl of one tothree carbons; R is more preferably methyl. The aromatic groups are morepreferably further substituted with sulfonyl, carboxy, alkyl of one to10 carbons or amino. Representative of groups for X and Y are

where, e.g., R is alkyl of 1 to 10 carbons, and R² is amino, nitro,sulfhydryl, hydroxy, alkoxy of one to three carbons, acetamino ormethyl.

The alkyl groups for X and Y may be branched or unbranched and includeup to ten carbons. Typical examples of alkyl groups for X and Y includemethyl, ethyl, n-propyl, isopcropcyl, n-butyl, sec-butyl-, tert-butyl,isobutyl, pentyl, hexyl, heptyl, octyl, nonyl and decyl. The alkylgroups may be in whole or in part in the form of rings such ascyclopentyl, cyclohexyl, cycloheptyl and cyclohexylmethyl. The cyclicgroups may be further substituted with alkyl or aryl groups.

Preferably, X and Y both contain aromatic groups. More preferably, X andY are both tosyl:

The groups for Z are the same as for X and Y or a fused aryl moiety.Fused rings for the Z position include naphthalene, phenanthrene,anthracene, indole, quinoline, isoquinoline, carbazole, benzimidazoleand benzofuran. Z is preferably an unfused group, more preferablybenezyl.

All groups for W, X, Y and Z may be further substituted with polarsubstituents such as NF₂, NO, NO₂, SH, SO₃H, OH, and CO₂H. These polargroups are capable of aiding solubility of the compounds.

Representative compounds include

3-Methylene-1,5-ditosyl-1,5,9-triazacyclododecane

5,9-Ditosyl-7-hydroxymethyl-1,5,9-tri-azabicyclo-[5,5,0]tridecane

5,9-Ditosyl-13-oxa-1,5,9-triazatricyclo[5,5,1^(1.7),1^(7,12)]-tetradecane

9-Benzyl-3-hydroxymethyl-1,5-ditosyl-1,5,9-triazacylododecane

9-Benzyl-3-chloromethyl-1,5-ditosyl-1,5,9-triazacyclododecane

3-Chloromethyl-1,5-ditosyl-1,5,9-triazacyclododecane

N,N-bis(3-toluenesulfonamidopropyl) toluenesulfonamide

1,5,9-Tritosyl-1,5,9-triazacyclododecane

3-Methylene-1,5,9-tritosyl-1,5,9-triazacyclododecane

3-Hydroxymethyl-1,5,9-tritosyl-1,5,9-triazacyclododecane

3-Chloromethyl-1,5,9-tritosyl-1,5,9-triazacylododecane

11-Methylene-1,5,9-triazabicyclo[7,3,3]pentadecane

1,5,9-Triazabicyclo[9,1,1]tridecane

9-Benzyl-3-keto-1,5-ditosyl-1,5,9-triazacyclododecane

9-Benzyl-3-methyl-1,5-ditosyl-1,5,9-triazacyclododecane

9-Benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane-9-oxide

9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane

9-Alkyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane

9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane epoxide

9-Benzyl-1-formyl-3-methylene-1,5,9-triazacyclododecane

9-Benzyl-1-formyl-3-methylene-5-tosyl-1,5,9-triazacyclododecane

9-Benzyl-3-methylene-1-tosyl-1,5,9-triazacyclododecane

9-Benzyl-3-methylene-1-acyl-5-tosyl-1,5,9-triazacyclododecane

9-(Ethoxycarbonyl)-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane

N-Benzylbis(3-benzenesulfonamidopropyl)amine

9-Benzyl-3-methylene-1,5-dibenzenesulfonyl-1,5,9-triazacyclododecane

N-Benzylbis[3-(N′-2-propenyltoluenesulfonamido) propyl]amine dihydrogensulfate

N-Benzyl-N-[3-(N′-2-methyl-2-propenyl-toluenesulfonamido)propyl]-N-(3-toluenesulfonamido-propyl)aminedihydrogen sulfate

N-Benzylbis[3-(N′-2-methyl-2-propenyltoluene-sulfonamido)propyl]aminedihydrogen sulfate.

The compounds of this invention possess valuable pharmacologicalproperties for both human and veterinary medicine. The compounds displayantiviral and antitumor effects and are useful particularly in theprevention and chemoprophylaxis of viral illnesses.

The compounds can also be used in vitro and employed in admixture withcarriers, germicides, fungicides, or soaps, etc., for use as antisepticssolutions and the like, particularly in conjunction with hospitalhousekeeping procedures to combat viruses such as herpes and HIV. Theyare also useful as intermediates in the production of other polyaminedrugs by the method of U.S. Pat. No. 5,021,409 and in the synthesis ofmetal chelating compounds, e.g., by methods of Bradshaw et al.,Aza-Crown Macrocycles, Wiley, New York, 1993.

The pharmacologically active compounds of this invention can beprocessed in accordance with conventional methods of pharmacy to producemedicinal agents for administration to patients, e.g., mammals includinghumans. The pharmacological compounds of the invention are generallyadministered to animals, including but not limited to mammals andavians; more preferably to mammals including humans, primates and aviansincluding poultry.

The compounds of this invention can be employed in admixture withconventional pharmaceutically acceptable diluents, excipients, carriersand other components such as vitamins to form pharmaceuticalcompositions.

Pharmaceutical compositions may be prepared by known principles forparenteral, enteral and topical application. Preferably, theadministration is parenteral.

Generally, the compounds of this invention are dispensed in unit dosageform comprising 10 to 1000 mg in a pharmaceutically acceptable carrierper unit dosage. The dosage of the compounds according to this inventiongenerally is about 0.1 to 110 mg/kg body weight/day preferably 0.1 to 20mg/kg/day when administered to patients, e.g., humans to treat viralinfections such as HIV.

Viruses share certain common characteristics; they consist of a nucleicacid genome which may be double-stranded DNA, single stranded DNA,single-strand positive RNA, single-strand negative RNA anddouble-stranded RNA. The nucleic acid is surrounded by protectiveprotein shell (capsid) and the protein shell may be enclosed in anenvelope which further includes a membrane.

The treatment of viral disease has been approached by inhibitingabsorption or penetration of virus into cells, inhibiting intracellularprocesses which lead to the synthesis of viral components, or inhibitionof release of newly synthesized virus from the infected cell. Theinhibition of one or more of these steps depends on the chemistry ormode of action of the virus.

The compounds of the invention have been shown to have antiviral effectagainst various viruses including retroviruses, particularly HIV whichresearchers believe to be a positive strand RNA virus. Effectiveness hasalso been shown against other viruses which infect humans includingcytomegalovirus and herpesvirus which are believed to be double strandDNA viruses, influenza virus which is believed to be a negative strandRNA virus, and also rous sarcoma virus which infects avians. Theinvention will be illustrated by the following non-limiting examples.

I.A. Synthesis of Compounds 1 and 2

The synthesis of N-benzylbis(3-toluene-sulfonamidopropyl)amine (compound2) and 9-benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclodecane (CADA)(compound 1) is shown in Scheme 1.

a. Bis(2-cyanoethyl)amine

Into a 1-L three-necked round-bottomed flask equipped with an additionfunnel, a dry-ice/acetone-cooled Dewar condenser, thermometer, andnitrogen inlet, was added 354 g (6.7 mol) of acrylonitrile. The additionfunnel was charged with 208 mL (3.2 mol) of concentrated ammoniumhydroxide, the apparatus was flushed with nitrogen and the acrylonitrilewas preheated to 70-75° C. by means of an 80° C. oil bath. The ammoniumhydroxide was added dropwise to the vigorously stirred reaction mixtureover a period of 2 h. Afterwards, the reaction mixture was stirredwithout external heating for 30 min., then heated to 70-75° C. by meansof a 75° C. oil bath for an additional 30 min. The excess acrylonitrileand most of the water were removed by rotary evaporation and the residuewas dried to constant weight under vacuum (0.5 mm Hg). The resultingyellow oil (387 g, 99%) was sufficiently pure to be used directly in thenext step, but can be distilled under vacuum, bp 186-190° C. (15 mm). ¹HNMR (CDCl₃) δ2.86 (t, J=6.6 Hz, 4 H), 2.44 (t, J=6.6 Hz, 4 H), 1.5 (br,1 H). ¹³C NMR (CDCl₃) δ118.4, 44.4, 18.8. IR (film, cm⁻¹) 3330 (s), 2920(s), 2850 (s), 2240 (s), 1440 (s), 1415 (s) 1360 (m), 1130 (s), 750(br).

b. N-Benzylbis(2-cyanoethyl)amine

A mixture of 50.0 g (0.406 mol) of bis(2-cyanoethyl)amine, 51.45 g(0.406 mol) of benzyl chloride, 1.0 g (6.7 mmol) of sodium iodide, 22.05g (0.208 mol) of sodium carbonate and 150 mL of acetonitrile was stirredmechanically and heated at reflux under nitrogen for 5 h. The cooledreaction mixture was filtered and the solids were washed withacetonitrile (3×50 mL). The combined filtrates were concentrated byrotary evaporation. A solution of the residue in 100 mL of CH₂Cl₂ waswashed with saturated aqueous Na₂S₂O₃ (2×20 mL) and saturated aqueousNaCl (2×50 mL). The combined aqueous layers were extracted with CH₂Cl₂(3×30 mL). The combined CH₂CO₂ solutions were dried over MgSO₄, filteredand concentrated by rotary evaporation. The residual solvent was removedunder vacuum (0.5 mm), yielding 70.4 g (89%) of product as a lightyellow oil. ¹H NMR (CDCl₃/TMS) δ 7.34 (m, 5 H), 3.70 (s, 2 H), 2.88 (t,J=6.8 Hz, 4 H), 2.44 (t, J=6.8 Hz, 4 H). ¹³C NMR (CDCl₃) δ137.5, 128.3,127.4, 118.4, 57.7, 49.1, 16.4. IR (film, cm⁻¹) 3070 (w), 3050 (m), 3020(s), 2930 (s), 2830 (s), 2240 (s), 1595 (m), 1575 (w), 1485 (s), 1445(s), 1415 (s), 1360 (br), 1250 (br), :1125 (s), 1070 (s), 1020 (s), 960(s), 730 (s), 690 (s).

c. N-benzylbis(3-aminopropyl)amine

A mixture of 43.3 g (0.203 mol) of N-benzylbis(2-cyanoethyl)amine, 7.8 gof a 50% aqueous slurry of Raney nickel and 90 mL of a 1.4 M solution ofNaOH in 95% ethanol was hydrogenated in a Parr apparatus for 48 h. Thecatalyst was removed by filtration and washed with 80 mL of 95% ethanol.The combined filtrates were concentrated by rotary evaporation. Asolution of the residue in 100 mL of hexane/chloroform (1:1, v/v) wasdried over Na₂SO₄, filtered and concentrated by rotary evaporation.Removal of solvent residues under vacuum (0.5 mm) afforded 40.3 g (90%)of product as a yellow oil. ¹H NMR (CDCl₃/TMS) δ7.31 (m, 5 H), 3.52 (s,2 H), 2.70 (t, J=6.8 Hz, 4 H), 2.45 (t, J=6.8 Hz, 4 H), 1.60 (quint.,J=6.8 Hz, 4 H), 1.26 (s, 4 H). ¹³C NMR (CDCl₃) δ139.5, 128.3, 127.7,126.3, 58.3, 50.9, 40.0, 30.6. IR (film, cm⁻¹) 3360 (br), 3270 (br),3070 (w), 3050 (w), 3020 (m), 2920 (br. s), 2850 (s), 2800 (s), 1600(s), 1485 (s), 1450 (s), 1360 (m), 1110 (br), 1070 (m), 1020 (m) 730(m), 690 (m).

d. N-Benzylbis(3-toluenesulfonamidopropyl)amine (compound 2)

A solution of p-toluenesulfonyl chloride (20 g, 105 mmol) in 50 mL ofCH₂Cl₂ was added dropwise with vigorous stirring over 2 h to a solutionof N-benzylbis(3-aminopropyl)amine (11.07 g, 50 mmol) and NaOH (4.4 g,110 mmol) in 30 mL of water. After stirring for an additional hour, theorganic layer was separated, washed with equal volume of brine, driedover MgSO₄, and concentrated by rotary evaporation. Attempts tocrystallize the crude oily product were not successful. All remainingsolvent was removed under high vacuum and the product (25.1 g, 95%) wasused without further purification. ¹H NMR (CDCl₃/TMS) δ7.70 (d, J=8.2Hz, 4 H, TsH^(2,6)), 7.28 (d, J=8.2 Hz, 4 H, TsH^(3,5)), 7.24 (m, 5 H,Ph), 5.80 (br, 2 H, NH, 3.43 (s, 2H, PhCH₂), 2.92 (t, J=6.2 Hz, 4 H,Ts-NCH₂), 2.45 (m, 4 H, Bn-N-CH₂), 2.42 (s, 6 H, TsCH₃), 1.64 (quint.,J=6.3 Hz, 4 H, NCH₂CH₂). ¹³C NMR (CDCl₃) δ143.1, 136.9, 129.6, 129.0,128.4, 127.3, 127.0, 58.6, 51.8, 42.2, 25.9, 21.4. IR (film, cm⁻) 3280,3050, 3020, 2940, 2850, 2810, 1595, 1490, 1450, 1320, 1150, 1085, 810,730, 690, 660.

The compound 9-benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclodecane(CADA) (compound 1) was synthesized in the following manner.

e. 9-Benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane (CADA)(compound 1)

Sodium hydride (3.6 g, 144 mmol, washed with hexane prior to use) wasadded under nitrogen to a solution of the product of procedure d.,N-benzylbis(3-toluenesufonamidopropyl)amine (26.5 g, 50 mmol) in 500 mLof DMF. The mixture was held at 80-100° C. for 1 h then cannulatedthrough a glass filter under N₂ into a 2-L three-necked round-bottomedflask equipped with a rubber septum, a thermometer, and an inlet for N₂.An additional 500 mL of DMF was used to insure complete transfer. Thesolution was stirred at 100° C. as a separate solution of3-chloro-2-chloromethyl-1-propene (6.25 g, 50 mmol) in 50 mL of DMF wasadded over 9 h by means of a syringe pump. Upon completion of theaddition, stirring at 100° C. under N₂ was continued an additional 12 h.The solvent was removed completely on a rotary evaporator using a hotwater bath. A solution of the residue in 150 mL of CHCl₃ was washed withwater, dried over MgSO₄ and concentrated by rotary evaporation. Asolution of the resulting sticky, yellow crude product in a minimumvolume of hot toluene was mixed with hexane to precipitateside-products. The supernatant solution was decanted and the residue wastriturated with hexane several times. The combined supernatants wereconcentrated by rotary evaporation. The resulting residue was dried invacuo and recrystallized from chloroform/ethanol, yielding CADA (16 g,55%) as a white solid, mp 156-158° C. NMR (CDCl₃/TMS) δ7.66 (d, J=8 Hz,4 H, TsH^(2,6)), 7.31 (d, J=8 Hz, 4 H, TSH^(3,5)), 7.20 (m, 5 H, Ph),5.23 (s, 2 H, C═CH₂), 3.84 (s, 4 H, allylic), 3.39 (s, 2 H, PhCH₂), 3.12(t, J=6.8 Hz, 4 H, Ts-NCH₂), 2.44 (s, 6 H, TsCH₃), 2.36 (t, J=6 Hz, 4 H,Bn-N-CH₂), 1.63 (quint., J=6 Hz, 4 H, NCH₂Cl₂). ¹³C NMR (CDCl₃) δ143.4,139.3, 138.3, 135.5, 129.7, 128.7, 128.1, 127.2, 126.9, 116.2, 59.0,51.0, 49.5, 44.0, 24.4, 21.5. IR (film, cm⁻) 3020, 2940, 2920, 2850,2800, 1600, 1490, 1450, 1330, 1150, 1080. Anal. calcd. for C₃₁H₃₉N₃S₂O₄:C, 64.00; H, 6.76; N, 7.22; S, 11.02. Found: C, 63.91; H, 6.65; N, 7.13;S, 11.04.

I.B. Synthesis of Compounds 210 and 211

210 N-Benzylbis(3-benzenesulfonamidopropyl)amine and 2119-Benzyl-3-methylene-1,5-dibenzenesulfonyl-1,5,9-triazacyclododecanewere synthesized as follows:

a. N-Benzylbis(3-benzenesulfonamidopropyl)amine (compound 210)

HCl Salt. Into a 1-L round-bottomed flask equipped with an additionfunnel, a stirring bar and a nitrogen inlet, were added 40.2 g(0.23 mol)of benzenesulfonyl chloride, 150 mL of ether and 175 mL of a saturatedaq. NaCl solution. The addition funnel was charged with a solution of25.4 g (0.12 mol) of N-benzylbis(3-aminopropyl)amine and 175 mL of 1.3 Naq. NaOH. The amine was added dropwise with vigorous stirring over 2 h.The mixture was poured into a separatory funnel and the aqueous portion(bottom layer) removed. The two organic layers were diluted with 100 mLof chloroform and the resulting solution concentrated by rotaryevaporation to a pale yellow oil. The residue was dissolved in 150 mL ofdichloromethane. A solution of 2 N aq. HCl (60 mL) and 60 mL ofsaturated aq. NaCl was added and the mixture was stirred vigourously for1 h. The layers were separated and the organic (lower) layer was washedwith a saturated aq. NaCl solution (3×50 mL) and concentrated by rotaryevaporation. The residue was recrystallized by bringing an ethanolsolution of the residue to a boil and adding ethyl acetate to induceprecipitation. The mixture was cooled to room temperature then to −25°C. for 2 weeks. Vacuum filtration of the precipitate gave 55.9 g (90%)of N-benzylbis(3-benzenesulfonamidopropyl)amine hydrochloride as a whitesolid. An analytical sample was prepared by drying at 65° C. (1 mm) for13 h, mp 135-136° C. ¹H NMR (DMSO-d₆) δ11.0 (br, 1 H, BnNH), 7.89 (t, 2H, J=5.7 Hz, SO₂NH), 7.79 (d, 4 H, J=7.2 Hz,SO₂ArH^(2,6)), 7.59(m, 8 H.SO₂ArH^(3,4,5), BnArH^(3,5)), 7.41 (m, 3 H, BnArH^(2,4,6)) 4.23 (d, 2 H,J=4.2 Hz, PhCH₂N), 2.93 (m, 4 H, ArSO₂NCH₂CH₂), 2.74 (m, 4 H,BnNCH₂CH₂), 1.88 (m, 4 H, NCH₂CH₂CH₂N), ¹³C NMR (DMSO-d₆) (140.3, 132.6,131.5, 129.8, 129.6, 129.4, 128.9, 126.6, 56.0, 49.4, 40.1, 23.3. IR(KBr, cm⁻¹) 3242 (br), 3073 (s), 2976 (m), 2855 (m), 2609 (s), 1470 (m),1446 (s), 1330 (s), 1163 (s), 1092 (s), 740 (s), 689 (s). Anal. calc.for C₂₅H₃₁N₃S₂O₄ HCl; C, 55.80; H, 5.99; N, 7.81; S, 11.92; Cl, 6.59.Found: C, 55.47; H, 6.13; N, 7.63; S, 11.63; Cl, 6.92%.

Free base. In a round-bottomed flask equipped with a magnetic stir bar,6.51 g (12.1 mmol) of N-benzylbis(3-benzenesulfonamidopropyl)aminehydrochloride, 40 mL of CH₂Cl₂, 15 mL of a solution of 1 N aq. NaOH and15 mL of saturated aq. NaCl were combined. After stirring vigorously for30 min, the layers were separated and the organic portion dried (MgSO₄).Removal of the drying agent by filtration, concentration of the filtrateby rotary evaporation and further drying under vacuum (0.4 mm, 45° C.)for 24 h afforded 5.96 g (98%) of N-benzylbis(3-benzenesulfonamidopropyl)amine as a thick colorless oil. TLC (silica): R_(f)=0.21, 1:1 (v/v) ethyl acetate:hexane. ¹H NMR (CDCl₃/TMS) δ7.82(d, 4 H, J=7 Hz,SO₂ArH^(2,6)), 7.51 (m, 6 H, SO₂ArH^(3,4,5)), 7.22 (m, 3H, BnArH^(2,4,6)), 7.16 (m, 2 H, BnArH^(3,5)), 5.9 (br, 2 H, SO₂NH) 3.40(s, 2 H, PhCH₂N), 2.92 (t, 4 H, SO₂NCH₂CH₂), 2.38 (t, 4 H, BnCH₂CH₂),1.62 (quint, 4 H, NCH₂CH₂CH₂N) ¹³ C NMR (CDCl₃) δ139.9, 138.2, 132.4,129.0, 128.4, 127.2, 126.9, 58.7, 51.8, 42.2, 26.1. IR (NaCl plate,cm⁻¹) 3277 (br), 3061 (w), 1446 (s), 1325 (s), 1160 (s), 1093 (s).

b. 9-Benzyl-3-methylene-1,5-dibenzenesulfonyl-1,5,9-triazacyclododecane(compound 211).

In a 500 mL three-necked round-bottomed flask equipped with a rubberseptum and a gas inlet, 0.92 g of a 60% (w/w) dispersion of sodiumhydride in mineral oil (0.55 g NaH, 23 mmol) was washed with hexane(3×15 mL) under N₂. A solution of 5.78 g (11.5 mmol) ofN-benzylbis(3-benzenesulfonamidopropyl)amine in 220 mL of DMF was addedslowly with stirring. The resulting clear solution was stirred under N₂while a solution of 1.44 g (11.5 mmol) of3-chloro-2-chloromethyl-l-propene in 8 mL of DMF was added dropwise over10 h. Stirring at room temperature under N₂ was continued for anadditional 24 h. The solvent was removed by rotary evaporationevaporation using a hot water bath. A solution of the residue in 50 mLof CHCl₃ was washed with water (3×30 mL) and saturated aq. NaCl (2×30mL) then dried (MgSO₄). Removal of the drying agent by filtration andconcentration of the filtrate by rotary evaporation gave the crudeproduct as a thick yellow oil. The oil was dissolved in 100 mL of 1:1(v/v) ethyl acetate/hexane and filtered through a pad of 16 g of basicalumina (Act 1, 80-200 mesh) washing with an additional 50 mL of 1:1ethyl acetate/hexane. Concentration of the combined filtrates undervacuum afforded 4.93 g (77 %) of9-benzyl-3-methylene-1,5-dibenzenesulfonyl-1,5,9-triazacyclododecane asa bubbly film that was converted to the hydrochloride salt.

HCl Salt. A solution of the crude product in 30 mL of CHCl₃ was stirredfor 30 m with 10 mL of 2 N aq. HCl and 10 mL of a saturated aq. NaClsolution. The organic layer was separated, washed with 20 mL ofsaturated aq. NaCl and concentrated by rotary evaporation. The residuewas recrystallized by bringing an ethanol solution to a boil and addingethyl acetate slowly until cloudiness occurs. After cooling to roomtemperature, the mixture was cooled to −25° C. for a period of 8 hgiving a white solid that was isolated by vacuum filtration andrecrystallized in a similar manner from acetone/ethyl acetate yielding3.03 g (45%) of pure compound 211 (hplc), mp 135-136° C. A samplesubmitted for elemental analysis was dried for 24 h at 78° C. (0.3 mm).¹H NMR (DMSO-d₆) δ11.6 (br, 1 H, BnNH), 7.79 (d, J=7.8 Hz, 4 H,SO₂ArH^(2,6)), 7.68 (m, 8 H, SO₂ArH^(3,4,5) BnH^(3,5)), 7.43 (m, 3 H,BnH^(2,4,6)), 5.37 (s, 2 H, C═CH₂), 4.30 (d, 2 H, PhCH₂N), 3.69 (s, 4 H,allylic), 3.09 (m, 8 H, SO₂NCH₂CH₂CH₂NBn), 1.95 (m, 4 H, NCH₂CH₂CH₂N).¹³C NMR (CDCl₃) δ141.5, 136.9, 133.4, 131.1, 130.4, 129.7, 129.4, 128.8,127.3, 118.8, 57.1, 51.8, 48.2, 46.3, 20.2. IR (KBr, cm⁻¹) 3060 (w),2928 (w), 2873 (w), 2445 (br), 1447 (s), 1335 (s), 1163 (s), 1090 (m),931 (m), 737 (s), 696 (m). UV (CH₃OH): λ_(max) (loge), 224 (4.2). FABMS:m/z 554 (M+H⁺, 100). Anal. calc. for C₂₉H₃₅N₃S₂O₄ HCl: C, 59.02; H,6.15; N, 7.12; S, 10.86; Cl, 6.01. Found: C, 59.39; H, 6.28; N, 7.21; S,11.34%

I.C. Synthesis of Compounds 214, 003 and 004

The synthesis of: 214N-Benzyl-N-[3-(N-2-methyl-2-propenyl-toluenesulfonamido)propyl]-N-(3-toluenesulfonamidopropyl)aminedihydrogen sulfate 003N-Benzylbis[3-(N′-2-methyl-2-propenyltoluenesulfonamido)propyl]aminedihydrogen sulfate 004N-Benzylbis[3-(N′-2-propenyltoluenesulfonamido)propyl]amine dihydrogensulfate is shown below:

a. N-Benzylbis[3-N′-2-propenyltoluenesulfonamido) propyl]aminedihydrogen sulfate (Compound 004)

In a 250 mL three-necked round bottom flask equipped with rubber septum,a thermometer, a condenser and a nitrogen inlet, a solution of 432 mg(0.79 mmol) of N-benzylbis(3-toluenesulfonamidopropyl)amine (compound 2)in 25 mL anhydrous DMF was added to 125 mg of 60% (w/w) of sodiumhydride in mineral oil (75 mg NaH, 1.88 mmol) that was previously washedwith hexane (3×15 mL). The mixture was stirred for an hour at roomtemperature while a solution of 0.30 mL (3.68 mmol) of 3-chloropropenein 4.0 mL of anhydrous DMF was added dropwise. Upon completion ofaddition, stirring at room temperature under N₂ was continued for anadditional 19 h. Vacuum filtration of the reaction mixture andconcentration of the filtrate in vacuo gave 0.5 g (100%) of the crudeproduct as a yellow oil. Free base: Preparative TLC of 202 mg of thecrude product using 40% EtOAc in hexane on silica gel GF plates (1000μm) yeilded 166 mg (82%) of the free base as a viscous, colorless oil.¹H-NMR (CDCl₃) δ7.66 (d, J=8.1 Hz, 4H, TsH^(2,6)), 7.27 (d, 4H,TsH^(3,5)), 7.24 (m, 5H, PhH), 5.61 (ddt, 2H, HC═CH₂), 5.11 (ddd, 4H,CH₂C═CHCH₂), 3.76 (d, J=6.3 Hz, 2H, NCH₂CH═CH₂), 3.44 (s, 2H, PhCH₂),3.09 (dd, 4H, TsNCH₂CH), 2.40 (s, 6H, NSO₂PhCH₃), 2.35 (t, J=6.9 Hz, 4H,BnNCH₂) 1.66 (quintet, J=7.2 Hz, 4H, NCH₂CH₂CH₂). ¹³C-NMR (CDCl)δ142.95, 139.36, 136.94, 133.27, 129.52, 128.65, 128.01, 127.00, 126.72,118.43, 58.3, 51.03, 50.70, 45.82, 25.98, 21.34. FT-IR (NaCl plate,cm⁻¹): 3064, 3027, 2924, 2802, 1920, 1643, 1598, 1494, 1453, 1418, 1337,1159, 1091, 1020, 928, 815, 736, 700, 660.

Compound 004: The purified free base (84 mg, 0.138 mmol) was dissolvedin ether and stirred vigorously at 0° C. as a cold solution of 0.43 MH₂SO₄ in ether (0.32 mL) was carefully added dropwise. The white solidthat precipitated was isolated by vacuum-filtration, washed with coldether and dried in vacuo to give 94 mg (96%) of Compound 004. ¹H-NMR(CDCl₃) δ7.66 (d, J=8.1 Hz, 4H, TsH^(2,6)), 7.27-7.42, (m, 5H, PhH),7.30 (d, 8.1Hz, 4H, TsH^(3,6)), 5.50 (ddd, 2H, HC═CH₂), 5.19 (dd, J=16.8Hz, 2H, trans-HC═CH₂), 5.16 (dd, J=9.9 Hz, 2H, cis HC═CH₂), 4.32 (s, 2H,PhCH₂) 3.72 (d, J=6.6 Hz, 4H, NCH₂CH═CH₂), 3.13 (m, 8H, BnCH₂CH₂), 2.41(s, 4H, TsNCH₂CH₂), 2.16 (s, 4H, NCH₂CH₂CH₂). UV (MeOH) :s(λ_(max))24,000 (232). FABMS: m/z 610 (MH³⁰, 100).

b. N-Benzylbis [3-N′-2-methyl-2-propenyltoluenesulfonamido)propyl)aminedihydrogen sulfate (Compound 003) andN-Benzyl-N-[3-(N-2-methyl-2-propenyltoluenesulfonamido)propyl]-N-(3-toluenesulfonamidopropyl)]aminedihydrogen sulfate (Compound 214)

A solution prepared from 50 mL of anhydrous DMF, 1.14 g (2.15 mmol) ofN-benzylbis(3-toluenesulfonamidopropyl)amine and 92 mg of 60% (w/w)sodium hydride in mineral oil (55 mg NaH, 2.30 mmol), which waspreviously washed with hexane (2×10 mL), was added over 2 h to a cold(0° C.) solution of 290 mg (3.23 mmol) of 3-chloro-2-methylpropene in 20mL anhydrous DMF. The reaction mixture was stirred at room temperatureunder N₂ for 22 h, then vacuum-filtered. The filtrate was concentratedin vacuo to give 1.49 of product mixture which was dissolved in CH₂Cl₂and separated by flash column chromatography using 230-400 mesh silicagel (Merck) and 45-50% EtOAc in hexane. The collected fractions wereanalyzed by TLC using 50% EtOAc in hexane. Compound 003 free base elutedfirst and was obtained as a colorless oil, 234 mg (19%). ¹H-NMR (CDCl₃)δ7.65 (d, J=8.1 Hz, 4H, TsH^(2,6)), 7.26 (d, J=7.8 Hz, 4H, TsH^(3,5)),7.22 (m, 5H, PhH), 4.84 (d, J=5.1 Hz, 4H, CH₂═CHCH₂), 3.63 (s, 4H,NCH₂CH═CH₂), 3.38 (s, 2H, PhCH₂), 3.04 (dd, J=7.8 Hz, 4H, TsNCH₂CH₂)2.40 (s, 6H, NSO₂PhCH₃), 2.26 (t, J=6.6 Hz, 4H, BnNCH₂), 1.70 (s, 3H,CH₃—CH═CH₂), 1.60 (quintet, J=7.8 Hz, 4H, NCH₂CH₂CH₂). ¹³C-NMR (CDCl₃)δ142.97, 140.94, 139.98, 136.98, 129.54, 128.70, 128.07, 127.10, 126.78,114.22, 58.14, 54.77, 51.14, 46.50, 25.79, 21.40, 19.75. FT-IR (NaClplate, cm⁻¹): 3028, 2944, 1813, 1658, 1598, 1494, 1454, 1336, 1160,1005, 918, 817, 699, 655. Anal. calc. for C₃₃H₄₇N₃O₄S₂: C, 65.90; H,7.43; N, 6.59; S, 10.05%. Found: C, 65.81; H, 7.31; N, 6.59; S, 10.14%.Compound 214 free base eluted next and was obtained as a colorless oil,422 mg (35%). ¹H-NMR (CDCl₃) δ7.67 (d, J=8.3 Hz, 4H, TsH^(2,6)), 7.25(m, 9H, PhH, TsH), 6.10 (br s, 1 H, TsNH), 4.87 (s, 2H, CH₂═CMeCH₂),3.64 (s, 4H, NCH₂CH═CH₂), 3.42 (s, 2H, PhCH₂), 3.06 (dd, 2H,TsRNCH₂CH₂), 2.94 (br dd, 2H, NHTsCH₂), 2.42 (s, 6H, NSO₂PhCH₃), 2.39(t, 2H, BnNCH₂CH₂CH₂NRTs), 2.33 (t, 2H, (BnNCH₂CH₂CH₂NHTs), 1.69 (s, 3H,CH₃—CH═CH₂), 1.69 (quintet, 2H, TsRNCH₂CH₂CH₂), 1.58 (quintet, 2H,TsHNCH₂CH₂CH₂). ¹³C-NMR (CDCl₃) δ143.15, 141.00, 138.61, 129.66, 129.56,128.98, 128.43, 127.22, 127.07, 114.47, 58.68, 54.96, 52.55, 51.25,46.61, 42.79, 25.76, 21.44, 19.18.

Compound 214: A solution of the purified free base (366 mg, 0.627 mmol)in 40 mL of ether was stirred vigorously at 0° C. as a cold solution of0.18 M H₂SO₄ in ether (2.0 ml,) was carefully added dropwise. The whitesolid that precipitated was isolated by vacuum-filtration, washed withcold ether and dried in vacuo to give 410 mg (96i) of Compound 214.¹H-NMR (CDCl₃ δ7.76 (d, J=8.4 Hz, 2H, TsH^(2,6)), 7.65 (d, J=8.4 Hz, 2H,TsH^(2,6)), 7.39-7.48 (m, 5H, PhH), 7.28(d, J=8.3 Hz, 2H, TsH^(3,5)),7.24 (d, J=7.8 Hz, 2H, TsH^(3,5)), 6.49 (br s, 2H, TsNH, HSO₄), 4.80(dd, 2H, CH₂═CMeCH₂), 4.28 (dd, 2H, TsRNCH₂CH₂), 3.52 (dd, 2H, PhCH₂),3.21 (br dd, 2H, TsHNCH₂CH₂), 3.01 (br dt, 6H, NCH₂CH═CH₂), 2.39 (t, 3H,NSO₂PhCH₃), 2.34 (s, 3H, NSO₂PhCH₃), 2.08 (quintet, 4H, NCH₂CH₂CH₂N),1.54 (s, 3H, CH₃—CH═CH₂). ¹³C-NMR (CDCl₃) δ143.57, 142.97, 140.06,137.12, 135.79, 131.42, 130.03, 129.81, 129.68, 129.39, 128.11, 127.332,127.15, 115.56, 57.042, 55.55, 51.122, 49.96, 45.80, 40.16, 24.08,23.24, 21.41, 21.38, 19.79. UV (MeOH) :s( max) 24,000 (232).

c. Compound 003

A solution of the purified free base (210 mg, 0.329 mmol) in 15 mL ofether was stirred vigorously at 0° C. as a cold solution 0.43 M H₂SO₄ inether (0.76 mL) was carefully added dropwise. The white solid thatprecipitated was isolated by vacuum-filtration, washed with cold etherand dried in vacuo to give 238 mg (98%) of Compound 003. ¹H-NMR (CDCl₃)δ7.68 (d, J=7.8 Hz, 4H, TsH^(2,6)), 7.53 (br d, J=4.2 Hz, 2H, PhH), 7.43(d, J=3.3 Hz, 3H, PhH), 7.30 (d, J=7.8 Hz, 4H, TsH^(3.5)), 4.86 (d,J=4.2 Hz, 4H), 4.28 (s, 2H, PhCH₂), 3.58 (s, 4H, NCH₂CH═CH₂), 3.09 (m,8H, TsNCH₂CH₂), 2.40 (s, 6H, NSO₂PhCH₃), 2.13 (quintet, 4H, NCH₂CH₂CH₂),1.61 (s, 3H, CH₃—CH═CH₂). ¹³C-NMR (CDCl₃) δ142.97, 140.94, 139.98,136.98, 129.54, 128.70, 128.07, 127.10, 126.78, 114.22, 58.14, 54.77,51.14, 46.50, 25.79, 21.40, 19.75. UV (MeOH):s (λ_(max)) 23,000 (232),Anal. calc. for C₃₅H₄₇N₃O₄S₂•H₂SO₄: C, 57.12; H, 6.71; N, 5.71; S,13.07%. Found: C, 57.97; H, 6.90; N, 5.77; S, 12.45%. FABMS: m/z 638(MH⁺, 100).

II. Synthesis of Compounds 12, 13, 14, 15, 16, 17

The synthesis of

12 3-Methylene-1,5-ditosyl-1,5,9-triazacyclododecane,

13 5,9-Ditosyl-7-hydroxymethyl-1,5,9-triazabicyclo-[5,5,0]tridecane,

14 5,9-Ditosyl-13-oxa-1,5,9-triazatricyclo[5,5,1^(1,7),1^(7,12)]-tetradecane,

15 9-Benzyl-3-hydroxymethyl-1,5-ditosyl-1,5,9-triazacyclododecane,

16 9-Benzyl-3-chloromethyl-1,5-ditosyl-1,5,9-triazacyclododecane, and

17 3-Chloromethyl-1,5-ditosyl-1,5,9-triazacyclododecane is shown inScheme 2.

a. The benzyl group of Compound 1 (CADA) was removed by reaction withalpha-chloroethyl chloroformate (ACE-Cl) giving the secondary amine(compound 12).

b. Intramolecular aminomercuration of 12 by reaction with mercuricacetate followed by alkaline reduction gave bicyclic alcohol 13 (53%)after separation of the product mixture by chromatography.

c. Oxidation of 12 with m-chloroperoxybenzozc acid gave isoxazolidine 14(20%), apparently via the nitrone produced by oxidation at nitrogenprior to epoxidation of the exocyclic double bond.

d. Functionalization of the alkene moiety was carried out viahydroboration of 1 with thexylborane, yielding alcohol 15 after alkalinehydrogen peroxide workup.

e. Chloro analogue 16 was then prepared by reaction of 15 withtriphenylphosphine and CCl₄, and debenzylation to 17 was performed byreaction of 16 with formic acid and 5% palladium-on-carbon.

III. Synthesis of Compounds 18, 19, 21, 22, 23, 24, and 26

The synthesis of

18 N,N-bis(3-toluenesulfonamidopropyl)toluenesulfonamide,

19 1,5,9-Tritosyl-1,5,9-triazacyclododecane,

21 11-Methylene-1,5,9-triazabicyclo[7,3,3]pentadecane,

22 3-Methylene-1,5,9-tritosyl-1,5,9-triazacyclododecane,

23 3-Hydroxymethyl-1,5,9-tritosyl-1,5,9-triazacyclododecane,

24 3-Chloromethyl-1,5,9-tritosyl-1,5,9-triazacyclododecane,

26 1,5,9-Triazabicyclo[9,1,1]tridecane is shown in Scheme 3.

a. The tritosylamide amide (compound 18) is prepared by tosylation ofN,N-bis(3-aminopropyl)amine (commercially available)

b. Cyclization of the disodium salt of 18 with propylene glycolditosylate gave macrocycle 19 (50%).

c. Macrocycle 19 was detosylated to 1,5,9-triazacyclododecane 20 (60%)by treatment with 98% sulfuric acid at 100° C.

d. Macrocyclic triamine 20 is commercially available, but is readilyprepared in multiple-gram quantities and the yield of 19 has beenimproved to 70%.

e. Reaction of 20 with 3-iodo-2-iodomethyl-1-propene gave bicyclicanalogue 21 in 60-70% yield.

f. Alkene 22(59%) was prepared by Richman-Atkins cyclization of 18 with3-chloro-2-chloromethyl-1-propene.

g. Another series of analogues was prepared from alkene 22.

By methods described previously for functionalization of compound 2(Scheme 1), hydroboration/oxidation of 22 gave alcohol 23, which wasthen converted to chloride 24 in greater than 90% yield overall. Thechloromethyl group of 25 survives the harsh conditions required fortosyl cleavage, yielding 25. Transannular cyclization was attempted bytreating this compound with potassium carbonate in isopropanol, but a59% yield of azetidine 26 was obtained along with a minor amount ofelimination product (27).

All of the analogues appearing in Schemes 1-3 can be synthesized inmultiple-gram quantities by these methods. Most can be purified byrecrystallization. Certain amines in these series occur as viscous oilsor glasses, but in many cases crystalline HCl salts can be obtained. TheHCl salts of 1 and 2 have also been prepared, with the expectation thatenhanced water solubility improves the ease of administration of thesedrugs.

IV. Synthesis of Compounds 41, 42, 43, 44, 45 and 46

Other symmetrical analogues

41 9-Benzyl-3-keto-1,5-ditosyl-1,5,9-triazacyclododecane

42 9-Benzyl-3-methyl-1,5-ditosyl-1,5,9-triazacyclododecane

43 9-Benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane-9-oxide

44 9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane

45 9-Alkyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane

46 9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane epoxide inthe family of triaza compounds to be used in the invention are shown inScheme 4.

Compounds 41 to 46 are closely related to CADA. They are designed tohave enhanced water solubility and to be capable of modification ofbiomolecules by electrostatic or hydrophobic interaction at the doublebond position for reversible binding of proteins, such as HIV capsidproteins, integrase or proviral DNA by SN² attack of an O, N or S at W.

Compounds 41, 42 and 43 are prepared in one step by oxidation orreduction of 1. Ketone 41 is prepared by ozonolysis of 1-HCl inmethanol, followed by reductive workup with dimethyl sulfide. Theexocyclic double bond is reduced to give 42 by catalytic hydrogenationunder conditions known for hydrogenation of disubstituted alkenes in thepresence of benzylamine Functionality.

Compound 41 has improved water solubility and 42 has similar polarity to1, but neither can undergo the S_(N)2′ covalent binding mechanismmentioned above.

Ketone 41 is capable of Schiff-base condensation with an amino group(e.g. of a lysine side-chain). Amine N-oxide 43 is prepared by reactionof 1 with hydrogen peroxide in methanol. The water solubility of thiscompound should be similar to that of 1•HCl so it can be used to testwhether the protonation equilibrium (1=1•CH⁺) is important to themechanism of action. Compound 12, previously prepared according toScheme 2, is an important analogue of 1 and an intermediate in thesynthesis of 44-46. Acylation to 44 (R═H, alkyl or aryl) replaces thebenzyl group with acyl groups of various sizes to test the steric andelectronic requirements for this site. If protonation of the nitrogenatom is important, then alkylation of 12 to 45 (R=methyl, ethyl,β-hydroxyethyl, isopropyl, etc.) independently tests the effects ofsteric bulk and polarity at this site. A series of analogues of type 45can be prepared in which R is a benzyl group bearing a polar substituent(OH, NO₂, SO₃H, CO₂H, NH₂ or NMe₂) to solubilize the drug. Finally,epoxidation of 44 with m-CPBA or Oxone will give 46 with enhancedpolarity and reactivity at the exocyclic site toward nucleophilicattack.

Synthesis of compound 43 was as follows:

9-Benzyl-3-methylene-1,5-ditosyl-1,5,9-triaza-9-N-oxide-cyclododecane(ARB 95-212) (Compound 43).

H₂O₂Method. In a 250 mL round-bottomed flask, 3.00 g (5.2 mmol) of CADAwas diluted with 100 mL of ethanol and 30 g of 30% H₂O₂. A magnetic stirbar was added, condenser attached and the mixture was heated by means ofan oil bath to reflux. After 5 h, the clear solution was allowed to cooland then concentrated by rotary evaporation. The residue was dilutedwith 50 mL of CHCl₃ and 30 mL of water and stirred briefly to dissolvethe contents. The organic layer was separated, washed with water (2×25mL) then with 25 mL of saturated aq. NaCl and dried (MgSO₄). Filtrationof the drying agent and concentration of the filtrate afforded a bubblyfilm. Further purification was performed by gravity columnchromatography with neutral alumina. (Act. 1, 60-325 mesh) using CHCl₃followed by 20:1 (v/v) chloroform:ethanol to elute Compound 43.Concentration of the fractions to a bubbly film followed by triturationwith acetone afforded 1.44 g (47i) of Compound 43 as a white solid, mp129-130° C.

Oxone Method. A mixture of 2.50 g (4.29 mmol) of CADA, 1.25 g (14.8mmol) of sodium hydrogen carbonate, 2.71 g (4.4 mmol) of Oxone, 25 mL ofwater and 75 mL of ethanol was stirred with a magnetic stir bar andheated (oil bath) to 45-50° C. for 24 h. After cooling to roomtemperature, the solid was removed by suction filtration and washed with40 mL of ethanol (CADA was recovered by washing the solid with CHCl₃ andconcentration of the chloroform filtrate). The combined ethanolfiltrates were concentrated and a CHCl₃ solution of the residue wasdried over MgSO₄. Removal of the drying agent by filtration andconcentration of the filtrate afforded 1.20 g (46%) of Compound 43 (¹HNMR) as a bubbly film. Purification by gravity column chromatographywith neutral alumina as described in the previous method afforded 0.35 gof Compound 43 as a white solid after trituration with acetone. A samplesubmitted for elemental analysis was dried at 400C (0.2 mm) for 4 h. TLC(alumina) : R_(f)=0.36, 20:1 (v/v) chloroform:ethanol. ¹H NMR(CDCl₃/TMS) δ7.65 (m, 2 H, BnArH^(3,5)), 7.62 (d, J=8.1 Hz, 4 H,TsH^(2,6)), 7.40 (m, 3 H, BnArH^(2,4,6)), 7.32 (d, J=8.1 Hz, 4 H,TsH³s), 5.33 (s, 2 H, C═CH₂), 4.29 (s, 2 H, PhCH₂), 3.70 (s, 4 H,allylic), 3.45 (t, 4 H, Bn-N(O)-CH₂), 3.10 (t, 4 H, Ts-NCH₂), 2.45 (s, 6H, TsCH₃), 2.10 (m, 4 H, NCH₂CH₂). ¹³C NMR (CDCl₃) δ144.0, 141.7, 133.7,132.4, 130.0, 129.8, 129.3, 128.3, 127.5, 118.8, 71.9, 61.2, 52.7, 48.8,22.0, 21.5. IR (KBr, cm⁻¹) 3063 (w), 3032 (w), 2951 (w), 2925 (w), 2867(w), 1598 (w), 1458 (m), 1339 (s), 1162 (s), 1090 (m), 1034 (w). UV(CH₃OH): λ_(max) (loge), 230 (4,4). FABMS: m/z 598 (M+H⁺, 41). Anal.calc. for C₃₁H₃₉N₃S₂O₅: C,62.29; H, 6.58; N, 7.03; S, 10.73. Found: C,61.92; H, 6,53; N, 7.24; S, 10.97%.

An analogue of compound 44 is compound 2139-(Ethyloxycarbonyl)-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane.

Compound 213 was prepared as follows:

9-(Ethyloxycarbonyl)-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane(compound 213). In an oven dried 250 mL round-bottoned flask, 8.20 g(14.1 mmol) of CADA was diluted with 65 mL of freshly distilled(P₂O₅)1,2-dichloroethane. A magnetic stir bar, reflux condenser with agas inlet and oil bath were added then the system purged with nitrogen.Ethyl chloroformate (1.70 mL; 17.8 mmol) was added by syringe and thesolution heated at reflux for 5 h. The solvent was removed using arotary evaporator with a 40° C. water bath. Dilution of the oily residuewith 60 mL of a solution of 2:3 (v/v) ethyl acetate:hexane caused awhite precipitate to form at room temperature after scratching theglass. The solid was filtered and washed with 3×30 mL of a solution of2:3 (v/v) ethyl acetate:hexane giving 6.63 g (83%) of Compound 213, mp125-126° C. (after drying at 78° C. and 0.9 mm for 19 h). TLC (silica):R_(f)=0.24, 1:1 (v/v) ethyl acetate:hexane. ¹H NMR (CDCl₃/TMS) δ7.68 (d,J=8.3 Hz, 4 H, SO₂ArH^(2,6)), 7.33 (d,J=8.3 Hz, 4 H, SO₂ArH^(3.5)), 5.21(s,2 H, C═CH₂), 4.07 (quart., J=7 Hz, 2 H, CO₂CH₂CH₃), 3.85 (s, 4 H,allylic), 3.33 (t, J=6.4 Hz, 4 H, EtO₂CNCH₂), 3.11 (m, 4 H, SO₂NCH₂),2.44 (s, 6 H, PhCH₃), 1.85 (m, 4 H, NCH₂CH₂CH₂N), 1.21 (t, J=7 Hz, 3 H,CO₂CH₂CH₃). ¹³C NMR (CDCl₃) δ157.0, 143.6, 139.4, 135.7, 129.8, 127.2,116.9, 61.2, 51.4, 45.5, 45.2, 28.0, 21.4, 14.7 IR (KBr, cm⁻¹) 2990 (w),2990 (w), 2948 (w), 1688 (s), 1598 (w), 1485 (m), 1424 (m), 1343 (s),1230 (s), 1154 (s), 1093 (s), 1027 (m), 903 (m), 818 (m), 788 (m).FABMS: ml/z 564 (M+H⁺, 100). Anal. calc. for C₂₇H₃₇N₃S₂O₆: C, 57.53; H,6.62; N, 7.45; S, 11.37. Found: C, 57.36; H, 6.54; N, 7.73; S, 11.74%.

V. Synthesis of Compounds 50, 51, 52 and 53

Another series of compounds is illustrated by formula II. Thesecompounds include

50 9-Benzyl-l-formyl-3-methylene-1,5,9-triazacyclododecane

51 9-Benzyl-1-formyl-3-methylene-5-tosyl-1,5,9-triazacyclododecane

52 9-Benzyl-3-methylene-1-tosyl-1,5,9-triazacyclododecane

53 e.g. 9-Benzyl-3-methylene-1-acyl-5-tosyl-1,5,9-triazacyclododecane

These compounds have three different substituents (X, Y and Z) on thethree nitrogen atoms of a 12-membered ring. The synthetic plan utilizesa series of reactions reported for selective, unsymmetrical substitutionof 1,5,9-triazacyclododecane. The intermediate is3-methylene-1,5,9-triazacyclododecane (27), which can be obtainedquantitatively by elimination of HCl from 25 Scheme 3. Reaction of 27with neat DMF dimethyl acetal gives 47 in high yield as shown in Scheme5. Monoalkylation with benzyl bromide will give 48 or 49 (or both). Bothof these salts are synthetically useful, but the completion of thesynthesis is illustrated with 48.

Alkaline hydrolysis will yield formamide 50, which is an unsymmetricalexample of formula 29, where X═H, Y=CHO and Z=benzyl. Tosylation of 50and alkaline hydrolysis will give 51 and 52 respectively. Compound 52 isa mono-detosylated analogue of 1 and the missing tosyl group can bereplaced with almost any alkyl, acyl, alkanesulfonyl or arenesulfonylgroup of interest (53, Y=various For example, Y=benzyl shows the effectof two protonation sites, while Y=p-bromobenzensulfonate shows theeffect of enhancing the sulfonamide leaving group ability. Candidatesfor improved water solubility include compounds of type 29 in whichZ=benzyl and X and Y are benzenesulfonyl groups bearing polarsubstituents, such as NH₂, OH or CO₂H.

VI. Synthesis of macrocyclic triamines of varying ring size: compounds55 and 60

Varied ring size and incorporation of amide functionality into the larering in macrocylic lactams are shown in Scheme 6.

Almost any linear triamine of type 54 (m, n=2-6, R=alkyl, aryl or acyl)is available by methods used for the synthesis of linear and macrocyclicpolyamines. Cyclization of these triamines with methyl acrylate orchloroacetyl chloride gives the corresponding macrocycles (55; a=2 or 1,respectively).

The acrylate cyclization is precedented by formation of thephenyl-substituted analogue of 55 (a=2, m=1, n=1, R═H) by reaction ofthe triamine with methyl cinnamate. Chloroacetyl chloride has been usedin the syntheses of numerous polyazamacrocycles by the so-calledcrablike cyclization, which is usually carried out in refluxingacetonitrile without resort to high-dilution conditions. The lowreactivity of the amide nitrogen atom of 55 enables selectivefunctionalization of the third nitrogen, affording 56 (Y=alkyl, acyl,alkanesulfonyl or arenesulfonyl). Both series of macrocyclic lactams (55and 56) can then be reduced to macrocyclic triamines under variousconditions for the reduction of amides to amines. The resultingcompounds are saturated analogues of type 29 bearing three different X,Y and Z groups, any of which could bear water-solubilizing substituents,and having ring sizes in the range of 9-18 carbon and nitrogen atoms.

VII. Synthesis of open-chain or non-macrocyclic analogues 58, 59, 60,61, 61, 63 and 64

Open-chain and non-macrocyclic analogues are shown in Scheme 7.

These analogues are synthesized from a series of l,n-alkanediamines viathe N-tosylphthalimide derivatives (57, n=2-7). Alkylation of thesodium, potassium or cesium salt of 57 with either2-chloromethyl-3-chloro-1-propene or 2-iodomethyl-3-iodo-l-propene willgive chain-extended intermediate 58 or 59. It is possible to control theproduct selectivity of the reaction by varying stoichiometry and orderof addition of the reactants. Intermediates of type 58 bear a secondleaving group (X), which will be displaced by various amine orsulfonamide nucleophiles, yielding 60 (Y═R′,SO₂R′ or SO₂Ar). Cleavage ofthe phthalimide protecting group by hydrazine will give primary amine61, which can be acylated or sulfonated to analogue series 62. The sameGabriel synthesis approach can be applied to 59 (n=2-8); anunsymmetrical series of compounds, differing only chain length relativeto 59, can be prepared by reaction of 58 with 57. The resulting seriesof compounds includes symmetrical and unsymmetrical open-chain analoguesof 1 containing the allylic tosylamide functionality and a total of 3-4nitrogen atoms.

VIII. Synthesis of bicyclic compounds 66, 67 and 68

An additional series of analogues consists of the bicyclic compounds34-36 shown below:

Examples of these bicyclic compounds and their syntheses are shown inScheme 8.

In bridged pyridine compound 66, corresponding to 34, the tertiary aminobasic site is replaced by a pyridine nitrogen atom and the aromaticbenzyl group is replaced by the aromatic pyridine ring. TheRichman-Atkins synthetic approach shown has been used successfully inthe preparation of many bridged pyridine host compounds. By analogy withknown cyclizations of 65 (n=1), reaction with2-chloromethyl-3-chloropropene or 2-iodomethyl-3-iodopropene underhigh-dilution conditions gives analogue 66 (n=1). The entire series inwhich n=1-6 is preparable by this method. In 35 and 36 the second ringis fused to two carbon atoms of CADA, leaving the center nitrogen atomfree for substituent Z. Two benzene fused examples of 35 and 36 are 67and 68, which are shown in Scheme 8 with their syntheses. The five stepsshown in each case parallel the 5-step synthesis of CADA (Scheme 1) andyields should be comparable. The fused benzene ring mimics the benzylgroup of CADA, leaving substituent Z variable. In all three cases(66-68) the fused aromatic ring can bear polar substituents enhancingwater solubility.

Other analogs such as those functionalized with polar groups forincreased solubility can be synthesized using the synthesis methodsdescribed above, coupled with various steps known in the art anddescribed, e.g., by T. J. Richman et al., Macrocyclic Polyamines:1,4,7,10,13,16-Hexaazacyclooctadecane in Organic Synthesis, Coll. Vol.VI, Noland, W. E., Ed., Wiley, New York, 1988, pp. 652-662; Bradshaw etal. Aza-Crown Macrocycles, Wiley, New York, 1993, chpt. IV.

Such polar groups include, for example, NO, NO₂, NH₂, OH, SH, OR, SR,COR, SO₂R, SOR, halide (F, Cl, Br, I) NHR, NR₂, HCO, COOH, COOR, C≡N,alkyl halide, etc. (R=alkyl of 1-10 carbons).

Antiviral activity was tested as follows:

EXAMPLE 1

In these tests, the IC₅₀ (50% inhibitory concentration) is theconcentration resulting in 50% cellular toxicity. EC₅₀ (median effectiveconcentration) is the compound concentration that reduces infection by50%. The effectiveness of a compound against a virus is expressed asTI₅₀ (median therapeutic index) which is determined by the ratioIC₅₀/EC₅₀.

Compounds of the invention were tested to assess the efficacy of theinhibitory effect of the compounds against HIV. The screen was carriedout in a soluble formazan assay by the tetrazolium method according toWeislow et al., J. Natl. Cancer Inst. 81:577-586, 1989.

The compound was dissolved in dimethyl sulfoxide (DMSO) (final conc.less than 0.25%) and then diluted 1:100 in cell culture medium. BecauseDMSO is toxic to cells at concentration above 1%, the concentration ofthe compound in DMSO should be at least 10 uM before dilution withaqueous solution. Various concentrations of the compound were testedagainst HIV-1 in CEM-IW cells. After six days incubation at 37° C. in a5% carbon dioxide atmosphere, viable cells were analyzed throughaddition of tetrazolium salt XTT followed by incubation to allowformazan color development. Cell viability was viewed microscopicallyand detected spectrophotometrically to quantitate formazan production.

In a test against HIV-1(6S)-AZT sensitive, the compound CADA(compound 1) was found active. The IC₅₀ index was greater than 5.00×10⁻⁵M; the EC₅₀ was 1.20×10⁻⁶ M; and the TI₅₀ (IC/EC) was greater than2.6×10⁺¹.

In a second test, the compound from another batch of CADA was confirmedactive in a primary screen. The IC₅₀ index was greater than 6.00×10⁻⁶M;the EC₅₀ was 2.40×10⁻⁶M; and the TI₅₀ was greater than 2.50×10°.

EXAMPLE 2

Testing was carried out as described in Example 1 against additional HIVstrains. The compound CADA was active against the following virusesusing the indicated cell lines:

IIIB strain of HIV-1/MT-4 cell line

HIV-2/CEM cell line

SIV/MT-4 cell line

AZT—resistant HIV-1/MT-2 and MT-4 cell lines

AZT—sensitive HIV-1/MT-4 cell line

A17, a strain of HIV-1 resistant to most of the non-nucleoside reversetranscriptase inhibitors/MT-2 and MT-4 cell lines,

WeJo, a clinical isolate of HIV-1/PBMCs (fresh human peripheral bloodmononuclear cells).

The compound was found to be active against these viruses with an EC₅₀of 2uM, a IC₅₀ greater than 30 μm and a TI₅₀ greater than 15. This meansthat good activity and low cell toxicity were observed in the micromolarrange of concentration of the compound.

EXAMPLE 3

Studies were carried out to determine the mechanism of action of thecompounds in the HIV life cycle. At least 16 steps in the HIV life cyclehave been identified as possible points for therapeutic intervention,yet all the anti-HIV drugs licensed in the U.S. so far (AZT, ddI andddC) are nucleoside inhibitors of HIV reverse transcriptase (RT).Another group of antivirals, the bicyclams, are believed to act at anearly stage in the retrovirus replicative cycle, apparently inhibitingviral uncoating. The action of other anti-HIVs such as the quinolines isunknown.

The studies were based on activity of the compound against HIV-1 inperipheral blood lymphocytes. The results with different solutions anddifferent batches of the compound confirm that the compound is active atsubmicromolar levels (EC₅₀) and has low cytotoxicity (IC₅₀). The resultsof the studies on the mechanism of reaction are summarized in the Tablebelow:

TABLE 1 Batch 1 A B Batch 2 PBL/HIV-1 Wejo EC₅₀ (μM) 0.55 0.16 0.65Infection IC₅₀ (μM) >100 >100 >100 Attachment p 24 based No InhibitionRT activity (rA.dT) ID₅₀ (μM) No Inhibition Time of Action Time LTR/gagNo Inhibition of Course proviral DNA synthesis Protease Activity ID₅₀(μM) 40 (rp HPLC) Mo-MO/HIV-1 Ba-L EC₅₀ (μM) 1.5 (fresh) Infection IC₅₀(μM) over 30 (high test) Latent Infection (U1/TNF) No inhibition

Preliminary mechanism of action studies showed that the compound CADAdoes not appear to inhibit HIV reverse transcriptase or HIV protease atthe concentrations determined to be effective in the in vitro assays. Itis believed that the CADA acts prior to the integration of virus intothe cellular genome, but it does not appear to inhibit virus attachmentor cell fusion.

It was concluded that the mechanism of action of the new compounds isdifferent than those of either of the two classes of AIDS drugscurrently in use or in clinical trials (reverse transcriptase andprotease inhibitors).

EXAMPLE 4

Additional studies on stability, solubility, formulation andpharmokinetics were carried out. Plasma elimination and urinary recoveryfrom mice following i.v. administeration were examined. Solubility ofthe compound in human, mouse, rat plasma and plasma ultrafiltrate wereexamined. Stability of the compound in human urine was determined.

The conclusions of these studies are as follows:

1. The compound is stable in mouse and rat plasma (t_(1/2) greater than200 hrs) and is stable in pH 4.0 buffer (t_(1/2) 139 hrs) and in humanplasma (t_(1/2) 126 hrs).

2. The compound is soluble in human plasma at 37° C. (1-2 ug/mL=2-3uM).

3. The compound can be formulated for animal studies at a concentrationof 1.2 uM in a 1:20 mixture of DMSO and normal saline (pH 2.7).

4. The compound is detectable in mouse plasma up to 2 hours after invivo intravenous administration.

EXAMPLE 5

Additional antiviral testing was carried out to assess the efficacy ofthe compounds of the invention against human cytomegalovirus (HCMV),herpes simplex virus (HSV), rous sarcoma virus (RSC) and influenza virus(FLU WSN).

Compounds tested were the HCl salts of the products compounds (1), (2)and (12) and non-salt compound (1). The results are summarized in thetable below

TABLE 2 IC₅₀, ug/mL Virus 2 · HCl CADA (1) 1 · HCl 12 · HCl HCMV5 >30, >50 PDR 3 HSV 11 10, 30 PDR 1.5 RSV 4 >30, >50 PDR 4 PDR FLUWSH >50 >30, >50 >50 12 PDR = poor dose response

The data in the table represents the concentration of compound causing50% protection against virus.

While not wishing to be bound by any one theory, one hypothesis is thatin the mechanism of action of the invention, the drug inhibitsretroviral uncoating by binding to a hydrophobic pocket on one of theHIV capsid proteins. A series of hydrophobic compounds, such asdisoxaril or Win 51711 are known to inhibit picornavirus replication bythis mechanism and various other drugs seem to neutralize rhinovirusesby binding to a specific hydrophobic pocket within the virion capsidprotein. The major HIV capsid protein (p24) is a potential target,although it has not been shown that such compounds can be as effectiveagainst an enveloped virus as they are against simple icosahedralviruses. Anti-influenza A drugs amantadine and rhimantidine inhibitvirus replication by blocking a proton channel associated with capsidprotein M2, thereby interfering with virus uncoating.

The bicyclams are the only reported inhibitors of HIV uncoating. CADAsuperficially resembles bicyclam as a macrocyclic polyamine. CADA'sbasic tertiary amino group should be protonated at physiologic pH andboth compounds could function as metal chelating agents. Thesesimilarities with bicyclams are only superficial, however, because thetoluenesulfonamide groups of 1 should render it a very weak metalchelator and 1 is much more lipophilic (hydrophobic) than bicyclamswhich should be polyprotonated a physiologic pH. Although a sulfonamide,CADA is cationic rather than anionic and does not belong to thepolyanionic sulfate class of anti-HIV agents that inhibit binding of thevirion to target cells.

Inhibition of the HIV enzyme integrase is a second hypotheticalmechanism of action consistent with the observation that CADA acts at anearly stage of HIV replication. Integrase controls the incorporation ofvirally transcribed DNA into the host genome, a key step in HIVreplication. Many drugs have been found to bind and inhibit HIV-1integrase in enzymatic in vitro experiments, but none of these compoundsprevents HIV replication in intact cells. It is possible that CADA isthe first therapeutically active drug operating by integrase inhibition.

Whether CADA is an uncoating inhibitor, an integrase inhibitor oroperates by another mechanism, its discovery leads to a new class ofdrugs complementing inhibitors of reverse transcriptase and protease,used individually or in combination therapy. CADA has a unique activityprofile and it operates at an early stage of HIV replication by amechanism not involving adhesion, fusion or reverse transcriptaseinhibition. The discovery shows the potential for a new approach to AIDSchemotherapy. It is likely that the synthesis of hundreds of analogueswill produce potent anti-HIV agents with improved solubility andbioavailability.

EXAMPLE 6

Additional antiviral testing was carried out to test the efficacy ofcompounds of the invention against human cytomegalovirus (HCMV) andanother herpesvirus, Variecella Zoster Virus(VZV). The compounds testedwere compound (1), the HCl salt of compound (1), Compounds 211, 43, 213and 214.

The results are summarized in the table below.

TABLE 3 Antiviral Screening Results for Herpesviruses HumanCytomegalovirus² Varicella Zoster Virus³ Compound EC₅₀ CC₅₀ SI EC₅₀ CC₅₀SI CADA (1) >34 133 <3.9 5.3 89 16.8 1 · HCl >160 >160 0 3.4 >160 >47.1211 >170 >170 0 91.7 >170 >1.8 43 >170 >170 0 24.3 >170 >7.0 213 3.0 16755.6 170 >180 >1.0 214 2.2 >150 >68 25.2 68.5 2.7 (20.5) (147) (7.1)¹Screening system: human foreskin fibroblast cells. EC₅₀ values areconcentrations (μM) required to inhibit viral cytopathogenicity by 50%.CC₅₀ values are concentrations (μM) required to inhibit cellproliferation by 50% (cytoxicity). SI (selectivity index) = CC₅₀/EC₅₀.Note: None of the compounds tested showed significant activity againstHerpes Simplex virus (HSV type 1 or 2) or Epstein Barr virus (EBV),except for Compound 213, which showed anti-EBV # activity (EC₅₀ 6.7 μM;CC₅₀ > 180 μM; SI > 26.9). ²Values not in parentheses are results of asemi-automated CPE-inhibition assay for human cytomegalovirus (HCMV).Values in parentheses are from plaque reduction assay. ³Results ofplaque reduction assay for VZV (Ellen strain).

EXAMPLE 7

Testing was carried out as described in Example 1 against additional HIVstrains MB48 and 2h. The results are summarized in the table below.

TABLE 4 ED₅₀ (μM)¹ Compound MB48/MT-2² MD48/HeLa³ 2h/PBMC⁴ CADA (1) 0.80.5 0.15 1 · HCl 0.2 0.1 0.15 211 1.9 1.5 0.5 43 1.8 3.5 0.5 004 2.6 3.50.5 ¹Concentrations required to inhibit viral cytopathogenicity by 50%for HIV strains and cell lines given. ²Reverse transcriptase (RT) assayin MT-2 cells. ³MAGI assay on HeLa-MAGIC cells (plaque inhibition).⁴Protein (p24) assay in PBMC cells with 2h, a primary pediatric clinicalisolate.

These results show antiviral activity in the submicromolar range.

EXAMPLE 8

The efficacy of intravenously administered CADA-HCl in rabbits duringocular Varicella Zaster Virus (VZV) infection was determined.Intravenous therapy with 5 mg/kg/day CADA-HCl for 11 days was comparedwith 20 mg/kg/day acyclovir (ACV) intravenous therapy.

Rabbits were bilaterally inoculated with an intrastromal injection of100 μL of McKrae strain VZV titer of 10⁵/ml. On day 2 to 3 postinoculation (PI) animals were evaluated and divided into equal groups.

Intravenous therapy of CADA-HCl, acyclovir and placebo saline wasinitiated immediately after animal grouping and continued through day 11PI. Two groups received CADA∩HCl intravenous therapy for 9 days, onegroup at a “low-dose” of 5 mg/kg/day and another group at a “high-dose”of 10 mg/kg/day. Other groups received similar IV therapy with ACV at 20mg/kg/day and with placebo.

As a result, VZV-induced corneal stromal and iris disease demonstrated apositive response to therapy with CADA-HCl and with ACV. CADA∩HCl atboth concentrations was effective in reducing the development of stromaldisease. By day 6 PI, acyclovir was intermediate between placebo and lowdose CADA-HCl.

Both CADA-HCl and ACV also showed a positive therapeutic response inreducing iris disease development. In efficacy against iris disease(uveitis) CADA-HCl and ACV both showed a positive effect. By day 5 PI,CADA∩HCl was more effective than acyclovir therapy in reducingdevelopment of iritis and in speeding resolution of VZV disease.

What is claimed is:
 1. A pharmaceutical composition havingpharmacological activity comprising a compound of formula I and apharmaceutically effective carrier:

wherein W is a bridge carbon which has a polar or non-polar side group,X and Y independently are an aromatic group, an alkyl group, a sulfonylgroup or a carbonyl group, said aromatic group is selected from thegroup consisting of Ar, Ar sulfonyl, Ar carboxy and Ar alkyl, where Aris an aromatic cyclic or aromatic heterocyclic ring having from five toseven members, said alkyl groups having from one to ten carbons, X and Yare not both an alkyl group so that at least one of X and Y is anaromatic group, a sulfonyl group or a carbonyl group, Z is a grouplisted for X and Y, a fused aryl moiety having from seven to ten carbonsor hydrogen, a, d and e independently are a number from zero to 10, cand b independently are a number from one to 10, and the compound iscyclic or acyclic and includes sufficient hydrogens for a stablemolecule.
 2. The pharmaceutical composition claim 1 wherein the polar ornon-polar side group for W is selected from the group consisting ofdouble-bonded carbon, double-bonded oxygen, hydroxyl, alkyl of one toabout 10 carbons, alkoxy of one to about 10 carbons, aryl of about sevento about 10 carbons, halogen, methyl halogen, methylene halide, epoxide,acyl, CH₂OH and hydrogen.
 3. The pharmaceutical composition of claim 1wherein the group for X and Y is further substituted with a hydrophilicgroup.
 4. The pharmaceutical composition of claim 1 wherein the groupfor X and Y is further substituted with NO, NO₂, NH₂, NHR, NHR₂, OH, OR,SH, SR, SOR, SO₂R, halo, C(halogen)₃,

where R is alkyl of C₁₋₁₀.
 5. The pharmaceutical composition of claim 1wherein X and Y are independently


6. The pharmaceutical composition claim 1 wherein c and b are three anda, d and e are independently zero or one.
 7. The pharmaceuticalcomposition claim 1 wherein W is ethene, X and Y are both tosyl and Z isbenzyl.
 8. The pharmaceutical composition claim 1 wherein the compoundis 3-Methylene-1,5-ditosyl-1,5,9-triazacyclododecane9-Benzyl-3-hydroxymethyl-1,5-ditosyl-1,5,9-triazacylododecane9-Benzyl-3-chloromethyl-1,5-ditosyl-1,5,9-triazacyclododecane3-Chloromethyl-1,5-ditosyl-1,5,9-triazacyclododecane1,5,9-Tritosyl-1,5,9-triazacyclododecane3-Methylene-1,5,9-tritosyl-1,5,9-triazacyclododecane3-Hydroxymethyl-1,5,9-tritosyl-1,5,9-triazacyclododecane3-Chloromethyl-1,5,9-tritosyl-1,5,9-triazacylododecane9-Benzyl-3-keto-1,5-ditosyl-1,5,9-triazacyclododecane9-Benzyl-3-methyl-1,5-ditosyl-1,5,9-triazacyclododecane9-Benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane-9-oxide9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane9-Alkyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane epoxide9-Benzyl-1-formyl-3-methylene-5-tosyl-1,5,9-triazacyclododecane9-Benzyl-3-methylene-1-tosyl-1,5,9-triazacyclododecane9-Benzyl-3-methylene-1-acyl-5-tosyl-1,5,9-triazacyclododecane9-(Ethoxycarbonyl)-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecaneN-Benzylbis(3-benzenesulfonamidopropyl)amine9-Benzyl-3-methylene-1,5-dibenzenesulfonyl-1,5,9-triazacyclododecaneN-Benzylbis[3-(N′-2-propenyltoluenesulfonamido)propyl]amine dihydrogensulfateN-Benzyl-N-[3-(N′-2-methyl-2-propenyl-toluenesulfonamido)propyl]-N-(3-toluenesulfonamido-propyl)aminedihydrogen sulfateN-Benzylbis[3-(N′-2-methyl-2-propenyltoluene-sulfonamido)propyl]aminedihydrogen sulfate; and the compound is in salt or non-salt form.
 9. Thepharmaceutical composition claim 1 wherein the compound is9-benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane,N-benzylbis(3-toluenesulfonamidopropyl)amine,3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane,9-(Ethoxycarbonyl)-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane,N-Benzylbis(3-benzenesulfonamidopropyl)amine,9-Benzyl-3-methylene-1,5-dibenzenesulfonyl-1,5,9-triazacyclododecane,N-Benzylbis[3-(N′-2-propenyltoluenesulfonamido) propyl]amine dihydrogensulfate,N-Benzyl-N-[3-(N′-2-methyl-2-propenyl-toluenesulfonamido)propyl]-N-(3-toluenesulfonamido-propyl)aminedihydrogen sulfate,N-Benzylbis(3-(N′-2-methyl-2-propenyltoluene-sulfonamido)propyllaminedihydrogen sulfate, or salts of these compounds.
 10. The pharmaceuticalcomposition of claim 1 wherein the pharmaceutical activity is antiviral.11. The pharmaceutical composition of claim 10 wherein the virus is aretrovirus, herpesvirus, influenza virus or rous sarcoma virus.
 12. Thepharmaceutical composition of claim 11 wherein the retrovirus is HIV.13. The pharmaceutical composition of claim 11 wherein the herpesvirusis Cytomegalovirus or Varicella zoster virus.
 14. A compound which is9-Benzyl-3-keto-1,5-ditosyl-1,5,9-triazacyclododecane9-Benzyl-3-methyl-1,5-ditosyl-1,5,9-triazacyclododecane9-Benzyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane-9-oxide9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane9-Alkyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane9-Acyl-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecane epoxide9-Benzyl-1-formyl-3-methylene-1,5,9-triazacyclododecane9-Benzyl-1-formyl-3-methylene-5-tosyl-1,5,9-triazacyclododecane9-Benzyl-3-methylene-1-tosyl-1,5,9-triazacyclododecane9-Benzyl-3-methylene-1-acyl-5-tosyl-1,5,9-triazacyclododecane9-(Ethoxycarbonyl)-3-methylene-1,5-ditosyl-1,5,9-triazacyclododecaneN-Benzylbis(3 -benzenesulfonamidopropyl)amine9-Benzyl-3-methylene-1,5-dibenzenesulfonyl-1,5,9-triazacyclododecaneN-Benzylbis[3-(N′-2-propenyltoluenesulfonamido)propyl]amine dihydrogensulfateN-Benzyl-N-[3-(N′-2-methyl-2-propenyltoluenesulfonamido)propyl]-N-(3-toluenesulfonamidopropyl)aminedihydrogen sulfate, orN-Benzylbis[3-(N′2-methyl-2-propenyltoluenesulfonamido)propyl]aminedihydrogen sulfate.
 15. The pharmaceutical composition of claim 1wherein the group for X and Y is further substituted with NH₂, NO, NO₂,SH, SO₃H, OH or CO₂H.